KR101742141B1 - - DHA EPA Fermented marine oil having high omega- essential unsaturated fatty acids DHA or EPA and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a culture concentrate of Rizopus sp. Microorganism which is fermented without any chemical process and no artificial emulsifier in marine oil (fish oil and algae oil collected from algae) to produce fermentation rich in free essential unsaturated fatty acids such as EPA and DHA Omega-3 essential unsaturated fatty acid DHA or EPA, which can maximize the physiological function of omega-3 such as blood triglyceride, blood pressure and atherosclerosis reduction, osteoporosis, ischemic dementia, etc., The present invention relates to a fermented marine oil having a high content and a method for producing the fermented marine oil, characterized in that a marine oil selected from the group consisting of fish oil, algae oil collected from algae and a mixture thereof is fermented with a culture concentrate of a microorganism of the genus Rizophus .

Description

[0001] The present invention relates to a fermented marine oil having a high content of DHA or EPA and a method for preparing the fermented marine oil having high omega-3 essential unsaturated fatty acids DHA or EPA,

The present invention relates to a fermented marine oil having a high content of free omega-3 essential unsaturated fatty acid docosahexaenoic acid (DHA) or eicosapentaenoic acid (EPA) and a preparation method thereof, and more particularly, By fermenting marine oils (bird oil extracted from fish oils and algae) with no chemical process and no artificial emulsifier with culture concentrates, fermented oils rich in free essential unsaturated fatty acids such as EPA and DHA are produced and their absorption Free omega-3 essential unsaturated fatty acid DHA or fermentation marine oil having a high EPA content, which can maximize the physiological function of omega-3 such as osteoporosis, ischemic dementia and the like, by reducing blood triglyceride, blood pressure and atherosclerosis, And a manufacturing method thereof.

Omega-3 essential fatty acid-containing oils have been known to be essential for normal growth and health since 1930, but scientific knowledge of their health benefits has been rapidly increasing and accumulating since the 1990s. Omega - 3 - containing oils grew at a rate of more than 50 - fold growth in domestic sales in 2009, compared to 2004. In 2011, Gyeonggi-do produced omega-3-containing oil, which is a health food containing more than 50 billion won of GHG. Sales of omega-3 essential fatty acid-containing oils in the Pacific region are estimated at 30% per year, and global market growth rates are as high as 15 to 20% per year. Consumption of omega-3 essential fatty acid-containing oils is expected to steadily increase in the future. do.

Omega-3 fatty acids, such as DHA and EPA, reduce the risk of coronary artery and cardiovascular disease by inhibiting platelet aggregation and lowering blood triglyceride levels. In addition, the effect of suppressing the rise in blood pressure and the relief of inflammation of rheumatoid arthritis was suggested. Especially, long-chain poly-unsaturated fatty acids such as alpha-linolenic acid (ALA), EPA and DHA have been investigated in various epidemiological studies and human and animal subjects, These omega-3 fatty acids are essential for the prevention and improvement of diverse circulatory system diseases such as vasodilating action, platelet aggregation inhibition, anti-ischemic action, increase of blood HDL-cholesterol and LDL-cholesterol reduction action, And it became more and more important as a fatty acid in the human body. In addition, these omega-3 fatty acids have been shown to be effective in relieving ADHD (Attention Deficit Hyperactivity Disorder), improving cognitive impairment due to aging, improving depression, and even more recently osteoporosis ) Have been reported to have a considerable effect on prevention and improvement.

In the body, carbohydrates and proteins undergo complex metabolism to produce unsaturated fatty acids containing saturated or double bonds. Essential fatty acids (EFAs) are essential for normal development and bio-health, but they are fatty acids that must be ingested in the form of food from the outside, since they are not synthesized in the human body. There are a number of bonds that are difficult to biosynthesis in the human body. For humans, ALA (alpha-linolenic acid, 18: 3n-3) and omega-6 fatty acid, LA (Linoleic acid, 18: 2n-6) are representative essential fatty acids. They are also called short chain-polyunsaturated fatty acids (SC-PUFA) because their carbon chain length is not very long, and ALA is more common in vegetable oils such as flaxseed oil. Conditionally essential fatty acids means fatty acids that can be biosynthesized in ALA or LA but must be replenished externally in vigorous development, disease, and aging conditions. Here, omega- 3 fatty acids such as EPA (20: 5n-3), DHA (22: 6n-3) and omega-6 fatty acids such as gamma-linolenic acid (18: 3n-6), dihomo-gamma-linolenic acid 20: 3n-6) and AA (arachidonic acid, 20: 4n-6). These fatty acids have relatively long carbon chains, and there are several double bonds, usually called LC-PUFA (long chain-polyunsaturated fatty acids). In general, the best way to increase the intake of omega-3 fatty acids (DHA and EPA) in the double is to increase the intake of fish, but it is not practical in practice. Therefore, it is recommended that these omega-3 fatty acid oils be supplemented to ordinary foods such as bread, milk, mayonnaise, pizza, yogurt, pasta, It is recommended in many advanced countries, and it is also being marketed actively as a formulation such as soft capsules. In particular, in 2004, the US Food and Drug Administration (EPA) found that omega-3 fatty acids in EPA and DHA were abundant in a variety of fish (tuna, salmon, mackerel, sardine, anchovy and herring) Low triglyceride levels, and significantly reduce the risk of heart disease. In the United States, many kinds of omega-3 fatty acid-containing foods are sold in the United States, from children's milk to processed meat products, and sales are steadily increasing every year. However, since the content of omega-3 essential unsaturated fatty acids is high in recent years, it has been suggested that fish oil which is generally consumed most is contaminated with heavy metals (mercury, lead, nickel, arsenic, cadmium etc.) and radioactivity. PCBs, furans, dioxins, etc. are also likely to be contaminated to a considerable extent. Among them, heavy metals are mostly associated with fish 's proteins, making it unlikely to accumulate in fat, the US Food and Drug Administration announced. Therefore, it was the key to digesting and absorbing the essential unsaturated fatty acids while consuming the minimum amount of fish oil.

DHA (Docosahexaenoic acid) is an unsaturated fatty acid having 22 carbon atoms (even number) and having 6 double bonds. This fatty acid plays an important role in infant brain and nerve cell membrane formation and maintenance, normal brain maturation and learning ability, and is effective in the prevention of cardiovascular diseases such as arteriosclerosis and myocardial infarction, and reduces accumulation of body fat.

EPA (Eicosapentaenoic acid) is an unsaturated fatty acid in which the number of carbon atoms is 20 (even number) and all double bonds are 5. It is a precursor of prostaglandin 13, which has antithrombotic effect and anti-arteriosclerotic effect, and is effective in preventing thromboembolic complications which is a cardiovascular disease.

The structural similarity between the two fatty acids is that the first double bond exists in the third carbon (omega-3) when counted from the omega carbon atom and that the existing double bond has a cis structure.

These fish, which are ingested as food, contain 98% or more of triacylglycerol or triglyceride (TG), a structure in which three molecules of fatty acid are ester-bonded to one molecule of glycerol, Or oil (hereinafter referred to as "oil"). This neutralization of the quality of the digestive tract is once the synthesis of the liver and the liver, emulsified by a large amount of secreted bile stored in the gallbladder, and then produced by the pancreas and secreted into the duodenum by pancreatic lipase And the ester bond between the glycerol and the fatty acid is hydrolyzed. As a result, free fatty acid and monoacylglycerol are mainly produced. It is known that triglycerides are in the form of monoacylglycerol, free fatty acids and glycerol before they are digested and spread to the intestines. They are known to diffuse and absorb into the small intestine epithelium. In general, the digestion and absorption of triglycerides is slower than other nutrients such as carbohydrates and proteins, and the absorption rate is also very low, and a large amount of bile secretion for digesting fat has been reported to adversely affect colon epithelial cells. Furthermore, the digestive enzymes that remove DHA or EPA, the essential unsaturated fatty acids of the present invention, from the glycerol skeletal molecule are less secretive than young and healthy people in those who are most in need of development of these essential unsaturated fatty acids and in the elderly There are aspects that are more difficult to digest and absorb.

There are three main types of omega-3 fatty acid-containing products currently on the market. The first form is a natural triglyceride (nTG) type that does not process fish oil at all. This is extracted and purified from fish, then sold in the form of triglycerides. As mentioned above, the bioavailability drops markedly and consumer consumption is gradually decreasing. The second type is a trans-esterification developed in the United States. After the oil is chemically reacted with ethanol to separate glycerol and fatty acid, the ethyl group is bonded to the carboxyl group of the free fatty acid, and the fatty acid ethyl ester molecule (FAEE) It will make. In particular, the EPA ethyl ester (EPA EE) and DHA ethyl ester (DHA EE) thus prepared are put into soft capsules and sold in the United States as general pharmaceutical products for improving blood triglyceride levels (Lovaza). However, the ethyl ester form of this type of omega-3 fatty acid is not covalently bound to the glycerol backbone molecule in vivo, but instead is covalently bound to the ethyl group, and pancreatic lipase must be operated again to remove the glycerol backbone. In this case, Which is about 50 times more difficult than removing fatty acids. Furthermore, after free fatty acids are absorbed by intestinal cells (Enterocyte) constituting the intestinal wall, they have to be changed into TG type in order to move in the blood, and there is no glycerol skeletal molecule, which makes it difficult to transfer blood. In order to overcome this problem, AstraZeneca in the UK recently developed a method for purifying only free omega-3 fatty acids already present in fish oil, but the production cost is too high, There is a risk of weakening. In addition, rTG (re-esterified TG) type in which the fatty acid of ethyl ester form is covalently bonded to glycerol is also sold. This is not only costly to manufacture but also has the drawback that there is no better absorption rate than nTG (natural TG) claims to those who developed them. Therefore, one single molecule of free fatty acid (FFA) type is most superior in terms of absorption rate, dosage efficiency, and bioavailability without covalent bonding with glycerol or ethyl group. For this purpose, a free fatty acid having a small amount in fish oil is purified It is not economically realistic. If FFA can be mass produced at low cost through the fermentation process, it can provide a wide range of applications in the food and pharmaceutical market.

To this end, instead of producing large quantities of Omega-3 fatty acids in pure FFA form, some preconditions must be met to meet the demanding needs of health-conscious consumers. In other words, it is necessary that the process is relatively eco-friendly because ethanol and other chemical catalysts are not used like the ester exchange method, and since the artificial emulsifier or other chemical catalysts or complex chemical processes are not used, It is necessary to be able to recover the free essential fatty acids produced and at the same time to be able to lower the unit cost because of the high recovery rate thereof. Finally, the bioavailability and the utilization rate of the thus- It should be superior to other formulations.

DISCLOSURE Technical Problem Accordingly, the present invention has been made keeping in mind the above problems occurring in the prior art, and an object of the present invention is to provide a method for efficiently producing free omega-3 essential unsaturated fatty acid having high dosing efficiency and bioavailability through fermentation of marine oil The content of free omega-3 essential fatty acid can be maximized by fermenting the marine oil under optimum conditions according to the present invention by using a culture concentrate of Rizophus genus microorganism instead of using the microorganism itself for fermenting marine oil, Thereby making it possible to manufacture a fermented marine oil capable of maximizing the effect of physiological effects such as improvement of digestion and bioavailability and improvement of blood circulation.

To this end, the present invention is characterized in that a marine oil is fermented by using a fermentation process, in particular, a culture concentrate of a microorganism of the genus Rizophus, instead of using the microorganism itself.

The fermentation marine oil according to the present invention is obtained by fermenting a marine oil selected from the group consisting of fish oil, algae oil extracted from algae and a mixture thereof with a culture concentrate of microorganisms of the genus Rizopus.

The culture concentrate of the Rizophus genus microorganism is inoculated with the microorganism of the genus Rizophus in an amount of 10 vol.% Relative to the medium volume and cultured under culture conditions at pH 5.5 to 6.5 and 25 to 35 ° C under aerobic conditions, It may be a precipitate obtained by salting out the supernatant obtained by centrifuging at 1 to 10 DEG C at 800 to 1200 rpm.

The medium may be a Potato Dextrose Broth (PDB) containing 2 to 10% by weight of potato starch, 0.5 to 3% by weight of dextrose, and water as a remainder. The culture using the PDB may preferably be performed at a pH of 5.1 + - 0.2 and under aerobic conditions, and the PDB may further comprise 0.1 to 10% by weight of oil based on the total weight of the PDB.

The culture time under the above culture conditions may be within a range of 80 to 150 hours.

The fermentation of the marine oil is carried out by adding a culture concentrate of the microorganism of Rizopus as a residual amount to 10 to 80% by weight of the marine oil and emulsifying it, Followed by fermentation under fermentation conditions comprising temperature, followed by purification of the fermentation marine oil.

The microorganism of the genus Rizophus may preferably be Rhizopus oryzae (microorganism resource center accession number KCTC6106).

The present invention also provides a method for producing a fermented marine oil by fermenting a marine oil, comprising the steps of: (1) selecting from the group consisting of fish oil, algae oil extracted from algae, and mixtures thereof; Mixing a culture concentrate of the Rizophus genus microorganism as a remainder in 10 to 80% by weight of the marine oil to be mixed and mixing to obtain a mixture; (2) an emulsifying step of emulsifying the mixture obtained in the mixing step; (3) After the emulsifying step, the emulsified mixture is agitated in a range of 50 to 500 rpm for 24 to 120 hours under a fermentation condition including a pH within a range of 5.0 to 8.0 and a temperature within a range of 25 to 45 DEG C A fermentation step for fermentation; And (4) a purification step of purifying the fermentation product obtained in the fermentation step.

The emulsification in the emulsification step may be carried out by stirring the mixture obtained in the mixing step for 5 to 20 minutes with a mixer and ultrasonication.

The purification in the purification step can be carried out by separating the fermentation product obtained in the fermentation step from the aqueous layer by centrifugation and filtering the oil layer.

The culture concentrate of the Rizophus genus microorganism is obtained by inoculating the microorganism of the genus Rizophus into the medium in an amount of 10% by volume based on the medium volume and culturing under culture conditions of pH 5.5 to 6.5 and 25 to 35 ° C under aerobic conditions, Followed by salting out the supernatant obtained by centrifugation at 800 to 1200 rpm at 1 to 10 < 0 > C.

The medium may be a Potato Dextrose Broth (PDB) containing 2 to 10% by weight of potato starch, 0.5 to 3% by weight of dextrose, and water as a remainder.

The culture time under the above culture conditions may be within a range of 80 to 150 hours.

The microorganism of the genus Rizophus may preferably be a Rythopus oryzae.

Further, the present invention provides a composition providing a wide spectrum in the food industry by containing the fermented marine oil according to the present invention as an active ingredient and greatly improving the taking efficiency and bioavailability of essential fatty acid omega-3.

The present invention also provides a fermented marine oil according to the present invention as an active ingredient to improve the effect of digestion and absorption of essential fatty acid omega-3, thereby lowering the triglyceride level in blood, Medicines, quasi-drugs or health foods.

Of course, the use of the fermented marine oil according to the present invention is not limited to the above functionality.

According to the present invention, in order to efficiently produce a free omega-3 essential unsaturated fatty acid having high dosing efficiency and high bioavailability through the fermentation process of marine oil, first, in order to ferment marine oil, By fermenting the marine oil at the optimal conditions according to the present invention by using a culture concentrate of microorganisms, the content of free omega-3 essential fatty acid is maximized, thereby greatly improving digestion and absorption and bioavailability, There is an effect of providing fermented marine oil which can maximize the effect.

FIG. 1 is a graph showing the production of free fatty acid of fermented oil prepared by first fermenting the oil collected from fishes and algae according to the present invention under optimal conditions selected in the present invention after HPLC analysis.
FIG. 2 is a graph showing the results of the physical and chemical analyzes of oils collected from fishes and algae according to the present invention, by performing physical emulsification (sonication and blending) without using an emulsifier and adding fermentation and emulsifier Tween-20 (0.2%, v / v) Which is obtained by fermenting a fermented oil produced by the fermentation of a fermented oil.
FIG. 3 is a graph showing the results obtained by blending oil extracted from fishes and algae according to the present invention with a blender for 10 minutes, and then sonifying the oil for a period of time to produce a physical emulsion and fermenting the fermented omega-3 oil Lt; RTI ID = 0.0 > HPLC < / RTI > analysis.
FIG. 4 is a graph showing the production of free fatty acid of fermented omega-3 oil prepared by secondary fermentation of oil extracted from fishes and algae according to the present invention after primary fermentation, after HPLC analysis.
FIG. 5 is a graph showing the production efficiency of free fatty acid by pH of a fermentation broth after HPLC analysis when fermenting oil extracted from fish and algae according to the present invention.
FIG. 6 is a graph showing the production efficiency of free fatty acids according to the fermentation temperature after the HPLC analysis according to the present invention when fermenting the oil collected from fishes and algae.
FIG. 7 is a graph showing the production efficiencies of free fatty acids per blending RPM (revolutions per minute) during the fermentation process to promote the physical emulsification and fermentation reaction of the whole fermentation broth when fermenting the oil collected from fishes and algae according to the present invention. Fig.
FIG. 8 is a graph showing changes in the concentration of DHA in the blood by time, after ingesting the samples collected before and after the fermentation of the fish and algae according to the present invention in a 24-hour fasted rat (6 weeks old) ). ≪ / RTI >
FIG. 9 is a graph showing changes in the blood concentration of EPA in time intervals after ingesting the oil collected from fishes and algae according to the present invention, before and after the fermentation and after fermentation for 24 hours in a rat (6 weeks old) chromatography).
FIG. 10 is a graph showing antimicrobial activity of Staphylococcus aureus of fermented oil obtained by fermenting oil extracted from fishes and algae according to the present invention. FIG.

Hereinafter, specific embodiments of the present invention will be described in detail with reference to the accompanying drawings.

The fermentation marine oil according to the present invention is characterized in that it is obtained by fermenting a marine oil selected from the group consisting of fish oil, algae oil collected from algae and a mixture thereof with a culture concentrate of microorganism of Rizophus.

That is, the present invention provides a fermented marine oil obtained by fermenting a culture concentrate of a microorganism of the genus Rizophus to an oil extracted from a marine oil, that is, fish and / or algae (algae).

For this purpose, the present invention uses a fermentation process. To this end, the microorganism used in the search is the strain that produces the most lipase that selectively hydrolyzes omega-3 fatty acids in the glycerol skeleton molecule. Instead of a conventional fermentation method in which a microorganism of the genus Rizophus is used as a microorganism, and in particular, a microorganism producing lipase is directly added to a marine oil and fermented, a culture concentrate of a microorganism of the genus Rhizopus, It was possible to minimize the possibility of microbial contamination of the finally produced fermented marine oil as well as minimizing the unique fermentation incurred in usual fermentation oils.

The fishes that can be used in the production of fish oil in the marine oil include mackerel, tuna, sardine, saury, salmon, mackerel and codfish. The algae are preferably Chlorella or Schizochytrium , It is to be understood that the present invention is not intended to be limited thereto and that the use of fish oil of other fish species rich in omega-3 fatty acids is also possible. The marine oil collected from the fish and / or algae contains a large amount of omega-3 fatty acids such as DHA and EPA, and is known to prevent vascular diseases such as hypertension, arteriosclerosis, heart disease, stroke, and inflammation of arthritis.

The oil taken from the fish and / or algae may be used as such or in the form of granules through sonification, vibration or physical stirring.

Table 1 below is data on the composition of various fatty acids of omega-3 oil (fish oil) used in the present invention. It is a result of analysis by AOAC method after separating all fatty acids from glycerol backbone molecules through chemical treatment, USA).

fatty acid How to measure result EPA AOCS 33% DHA AOCS 22% Other omega-3 fatty acids
(18: 3, 18: 4, 20: 4, 21: 4, 21: 5, 22: 5) AOCS 10.5% Other fatty acids AOCS 32%

The culture concentrate of the Rizophus genus microorganism is obtained by inoculating the microorganism of the genus Rizophus into the medium in an amount of 10% by volume based on the medium volume and culturing under culture conditions of pH 5.5 to 6.5 and 25 to 35 ° C under aerobic conditions, The precipitate obtained by salting out the supernatant obtained by centrifuging at 800 to 1200 rpm at 1 to 10 DEG C and using the culture concentrate of the Ricofus genus microorganism to directly ferment the microorganism using the concentrate, 3 fatty acid can be greatly increased.

At this time, the medium may be potato starch, PDB (Potato Dextrose Broth) containing 2 to 10% by weight of potato starch, 0.5 to 3% by weight of dextrose and water as a remainder. The culture using the PDB may preferably be performed at a pH of 5.1 + - 0.2 and under aerobic conditions, and the PDB may further comprise 0.1 to 10% by weight of oil based on the total weight of the PDB.

Particularly, according to the present invention, it was confirmed by the present inventors that the fermentation culture time of the cultured concentrate of the microorganism of the genus Rizophus is the best when the microorganism is cultured within a range of 80 to 150 hours.

The Rhizopus sp. Microorganism is one of the fungi of the fungus Mucorales. It is a branch of the stolon that spreads sideways and extends to the point where it touches the medium. node, and one to several sporangia and rhizoids. Sporangia are large, usually black. The columella is hemispheric, mycelium occurs in large quantities, and it also grows long to fill the Petri dish lid. In essence, two spousal cysts contact and fuse to form junction spores. Soil, fruit, etc., and rarely human, animal, or some species of plants are pathogenic. It has a strong starch sugar strength and a very small amount, but it has alcohol fermentation ability and proteolytic ability and also has a strong ability to produce organic acids such as lactic acid and fumaric acid. For example, it is used for the brewing of Chinese sake and the production of rice wine in Southeast Asia. R. delemar and R. japonicus are important as amylose for the production of alcohol. R. nigricans is also known to occur in fruits such as peaches and strawberries, cereals, and bread, and also causes sweet potato disease, but it is also known to have a strong fumaric acid-producing ability.

The culture concentrate of the microorganism of the genus Rizophus contains a lipase, and the lipase means an enzyme which decomposes into glycerol and fatty acid.

The fermentation of the marine oil is carried out by adding a culture concentrate of the microorganism of Rizopus as a residual amount to 10 to 80% by weight of the marine oil and emulsifying it, The fermentation can be carried out by fermenting the fermentation marine oil after fermentation under a fermentation condition including temperature, and the microorganism of Rizophus may preferably be a Rythopus orienting agent.

The above-mentioned fermentation marine oil obtained according to the above has a high content of essential unsaturated fatty acid which is liberated before fermentation. The free essential unsaturated fatty acids include DHA or EPA. DHA plays an important role in brain and nerve cell membrane formation and maintenance, normal brain maturation and learning ability, and is effective in preventing cardiovascular diseases such as arteriosclerosis, myocardial infarction, and reducing accumulation of body fat. EPA is a precursor to prostaglandin 13 that has anti-thrombotic effects and anti-arteriosclerotic effects and is effective in preventing thromboembolic complications from cardiovascular disease.

Therefore, the fermented omega-3 oil of the present invention has increased absorption of omega-3 essential unsaturated fatty acid in the glycerol skeleton molecule and thus increased the absorption rate in the small intestine so that the functional effect such as blood circulation improvement is less than that of the nTG type oil Is expected to be even better. In addition, as the content of the free essential unsaturated fatty acid increases, the stickiness is reduced compared to the omega-3 oil before fermentation, and the efficiency and the bioavailability are increased. In the past, omega-3 fatty acid DHA or EPA was taken intact or dissolved in ethyl alcohol to be taken or purified. However, in this case, a chemical process such as the use of an emulsifier may be added to the body to deteriorate the human body. This is a low problem. On the other hand, by pre-fermenting omega-3 fatty acid DHA or EPA, the present invention can reduce cost and time compared with chemical processes, accelerate digestion and absorption, and thus obtain the same effect even when taking a small amount, Omega-3 Fatty Acids By allowing covalent bonds in DHA or EPA to be easily broken without the use of an emulsifier, it is easy to separate in vivo, and the odor can be removed excellently.

The present invention also provides a method for producing a fermented marine oil by fermenting a marine oil, comprising the steps of: (1) selecting from the group consisting of fish oil, algae oil extracted from algae, and mixtures thereof; Mixing a culture concentrate of the Rizophus genus microorganism as a remainder in 10 to 80% by weight of the marine oil to be mixed and mixing to obtain a mixture; (2) an emulsifying step of emulsifying the mixture obtained in the mixing step; (3) After the emulsifying step, the emulsified mixture is agitated in a range of 50 to 500 rpm for 24 to 120 hours under a fermentation condition including a pH within a range of 5.0 to 8.0 and a temperature within a range of 25 to 45 DEG C A fermentation step for fermentation; And (4) a purification step of purifying the fermentation product obtained in the fermentation step.

The mixing step of (1) above is carried out by adding a culture concentrate of the microorganism of the genus Rizophus as a remaining amount to 10 to 80% by weight of a marine oil selected from the group consisting of fish oil, bird oil extracted from algae and a mixture thereof, , Wherein the marine oil is preferably preheated to a temperature within the range of 10 to 40 占 폚.

The emulsification step (2) comprises emulsifying the mixture obtained in the mixing step. The emulsification in the emulsification step is preferably carried out by stirring the mixture obtained in the mixing step for 5 to 20 minutes with a mixer Followed by sonication. The ultrasonic wave treatment may be performed by applying ultrasonic waves to the mixture. The application of such ultrasonic waves can be performed by purchasing a commercially available ultrasonic wave generator, which is commercially available by a manufacturer of domestic and foreign ultrasonic wave generators So long as it can be suitably used according to the manufacturer's instructions.

Wherein the fermentation step (3) comprises, after the emulsification step, heating the emulsified mixture at a pH within the range of 5.0 to 8.0, at a temperature within the range of 25 to 45 DEG C for 24 to 120 hours at a temperature of 50 to 500 rpm And the mixture is fermented while stirring.

The purification step (4) consists in purifying the fermentation product obtained in the fermentation step, and preferably the purification step separates the fermentation product obtained in the fermentation step from the aqueous layer from the aqueous layer by centrifugation , And filtering it. Here, the centrifugation preferably separates the fermentation product from the oil layer and the aqueous layer by centrifugation within the range of 8,000 to 12,000 rpm, removes the aqueous layer from the oil layer, and then filtrates the remaining oil layer ≪ / RTI > Also, here, the filtration can be preferably ultrafiltration, which is one of the membrane separation techniques, in which molecular-level filtration, which separates large solute molecules from small solute molecules or solvent molecules, . The membrane separation technique is usually classified into filtration, microfiltration, ultrafiltration, and reverse osmosis. Depending on the size of the material to be separated, ordinary filtration is up to 10 μm, microfiltration is up to 30 nm, ultrafiltration is up to 2 nm, and reverse filtration is less than that. By using a hole-free membrane made of a synthetic polymer such as cellulose acetate, the small molecule is infiltrated by pressurization. The polymer solution is used for concentration, desalination, fractionation, etc. Those who are familiar with the art can purchase commercially available products supplied by leading domestic and foreign ultrafiltration manufacturers and use them appropriately in accordance with the manufacturer's instructions As is well known in the art.

In the above method, the oil is preferably an oil extracted from algae such as mackerel, tuna, sardine, saury, salmon, mackerel or cod and fish such as Chlorella or Schizochytrium , DHA, EPA, and other essential omega-3 fatty acids.

In particular, the culture concentrate of the Rizophus genus microorganism is obtained by inoculating the microorganism of the genus Rizophus into the medium in an amount of 10% by volume based on the culture medium and culturing under the culture conditions of pH 5.5 to 6.5 and 25 to 35 ° C under aerobic conditions Followed by salting out the supernatant obtained by centrifugation at 1 to 10 ° C at 800 to 1200 rpm. The microorganism of the genus Rizophus is cultivated in the medium for a predetermined time so that lipase can be produced in high frequency, and the culture is centrifuged Or filtration to remove the cells completely and ammonium sulfate is slowly added to the remaining filtrate to cause enzyme precipitation and centrifugation again to obtain a high-concentration culture concentrate. The culture concentrate is sodium phosphate (sodium phosphate buffer solution (50 mM, pH 5 to 8), dialyzed into the same buffer solution, and then used.

The medium may be a Potato Dextrose Broth (PDB) containing 2 to 10% by weight of potato starch, 0.5 to 3% by weight of dextrose, and water as a remainder.

The culture time under the above culture conditions may be within a range of 80 to 150 hours.

The microorganism of the genus Rizophus may preferably be a Rythopus oryzae.

The present invention also provides a composition which provides a broad spectrum in the pharmaceutical or food industry by improving the efficiency of taking omega-3 essential fatty acids containing the fermented marine oil according to the present invention as an active ingredient, bioavailability and stability do.

When the fermented marine oil according to the present invention is used for medicines or foods, the fermented omega-3 oil rich in EPA and DHA rich in free essential unsaturated fatty acids can be taken to improve the dosage efficiency and bioavailability, Improvement of growth for ischemic dementia, antiinflammation, blood cholesterol level and triglyceride level can be improved for smooth circulation.

Specifically, the present invention relates to a method for improving the blood circulation improving the efficiency, bioavailability and stability of the essential fatty acid omega-3 containing the fermented marine oil according to the present invention as an active ingredient, or improving blood circulation, blood cholesterol, blood triglyceride, Reduction of sclerosis, or anti-inflammatory pharmaceutical composition, health food or quasi-drug.

The present invention also provides a pharmaceutical composition for antimicrobial use containing the fermented marine oil according to the present invention as an active ingredient, a health food or a quasi-drug.

Hereinafter, preferred embodiments and comparative examples of the present invention will be described.

The following examples are intended to illustrate the invention and should not be construed as limiting the scope of the invention.

Experimental Example 1 Comparative Experiment of Lipase Activity by Microorganism

Experiments were performed to compare the lipase activity of the culture concentrate of the Rizophus genus microorganism according to the present invention with other microorganisms.

As the control microorganisms, Burkholderia multivorans contains 1 wt% peptone, 0.5 wt% yeast extract, 0.02 wt% magnesium sulfate, 0.1 wt% potassium phosphate dibasic, 1 Pseudomonas fluorescence is a nutrient medium (nutrient medium) which is a type of bacterial medium which generally uses an extract of fish or meat meal, and mainly contains byproduct organic matter 0.5% by weight of peptone, 0.5% by weight of yeast extract, 0.8% by weight of salt, etc. may be added to 0.3% by weight of commercially available soup or especially for bacteriological extract to adjust the pH to about 7 1% olive oil was used for the culture medium, and Candida rugosa was used as a culture medium for YMB (1.5 g of yeast extract, 1.5 g of malt extract 1.5 g, peptone (2.5 g), dextrose (5.0 g), and distilled water (500 ml)) was added with olive oil to 1 wt% of the total amount of the medium. The medium of Pseudozima sp. Contains 0.01% glucose, 0.1% olive oil, 0.003% yeast extract, 0.003% malt extract, 0.003% peptone, 0.003% soybean flour, 0.02% ammonium sulfate , 0.001 wt% potassium phosphate, 0.005 wt% magnesium sulfate, 0.001 wt% calcium chloride, and 0.0001 wt% sodium chloride were used.

In order to measure the activity of lipase, a coloring method using a saponification reaction was used. The microorganisms were cultured, centrifuged at 12,000 rpm, and the supernatant was used as a crude enzyme solution. The substrate was incubated at 37 ° C for 30 minutes with 1% polyvinyl alcohol containing 10% olive oil, and the resulting free essential fatty acid was treated with copper-acetate- It is defined as μmol of the fatty acid generated in 1 minute of color development when copper is reacted with pyridine (Cu-acetate-pyridine).

The highest pH and time of lipase activity according to each microorganism are shown in Table 2 below.

pH time Lipase activity
(U / ml) Rhizopus oryzae 6.0 120 4.3 Burke Alderia Multi Bowl
( Burkholderia multivorans ) 7.8 36 0.8 Pseudomonas fluorescens
( Pseudomonas fluorescence ) 7.0 32 1.2 Candida rugosa 7.2 100 1.6 Pseudozima sp. 7.0 24 2.8

As shown in Table 2, it was confirmed that the Ricofus orientalis used in the present invention exhibited the highest lipase activity by culturing.

Experimental Example 2 Comparative Experiment of Lipase Activity by Culture Time

The lipase activity was measured according to the incubation time of Rissofus oryzae exhibiting the highest lipase activity in Experimental Example 1 above. The experiment was carried out under the same culture conditions as in Experimental Example 1, except that the incubation time was different, and the results are shown in Table 3 below.

Strain Incubation time Lipase activity
(U / ml)
Rhizopus oryzae
90 hours 4.15 120 hours 4.30 140 hours 4.22

As shown in Table 3, in the case of Rizoffus orientalis, the lipase activity showed the highest activity at 120 hours of culture time, and then gradually decreased with time.

≪ Experimental Example 3 > Concentration of lipase produced from the culture solution and measurement of activity of lipase

As a microorganism, Rissofus oryzae was cultured and centrifuged at 8000 rpm. Each of the cultures was treated with ammonium sulfate at a concentration of 20 to 100% by weight at 4 캜 to prepare a protein containing lipase And concentrated to give a concentrate. Then, the supernatant was removed by centrifugation at 4 ° C and 12,000 rpm for 20 minutes, and the precipitate was dissolved in the same volume of 0.05 M phosphate buffer solution (pH 7.0), and then dialyzed against the same buffer solution (4 ° C.) A concentrated concentrate was obtained.

The activity of lipase according to the saturated concentration of ammonium sulfate used for lipase precipitation is shown in Table 4 below.

Ammonium sulfate concentration The lipase activity (U / ml) 0 to 20% 3.54 20 to 40% 10.22 40 to 60% 14.84 60 to 80% 15.33 80 to 100% 15.48

As can be seen from the above table, the activity of lipase was highest when the saturation concentration of ammonium sulfate used in the lipase precipitation was 80 to 100%. However, considering the economical aspect, the concentration of ammonium sulfate is suitably from 60 to 80%.

PREPARATION EXAMPLE 1 Preparation of a Culture Concentrate of Risofussa Microorganism

Rhizopus oryzae as a microorganism belonging to the genus Rhizopus was cultivated in PDB (Potato Dextrose Broth) containing 4% by weight of potato starch, 1% by weight of dextrose, 1% by weight of olive oil and water as a remainder, And 10% by volume relative to the culture medium, culturing under culture conditions of pH 6.0 and 30 ° C under aerobic conditions, followed by centrifugation at 1 to 10 ° C at 800 to 1200 rpm to obtain a precipitate obtained by salting out the supernatant obtained , Which was used as a culture concentrate of microorganisms of the genus Rizophus.

<Example 1 and Controls 1 and 2> Fermentation performance comparison between microorganisms (Pseudomonas sp. And Rythopus orientalis) and Rissofus orientalis culture concentrate

(Example 1) of the Rizophus sp. Microorganism ( Rizophus orientalis ) according to the present invention of Preparation Example 1 and Pseudozima sp. Microorganism (Control 1: Pseudozima sp. ) As a microorganism which was not concentrated in culture as a control group . The fermentation of fish oil collected from sardine as a marine oil was tested for differences in fatty acid content using fermented fish oil (control 2). However, the culture concentrate of Rissofus oryzae was prepared by mixing the product obtained in Preparation Example 1 with a sodium phosphate buffer (50 mM) at pH 7.0.

The microorganisms Pseudojima and Rythopus orientus as fermented by the microorganisms themselves as control were inoculated into the PDB medium in an amount of 10 vol% relative to the medium volume, And 50% by weight of fish oil obtained from sardine as marine oil at a ratio of 50% by weight of each of the Rissofus oryzae culture concentrates, fermented at 37 ° C for 72 hours at a rotation speed of 300 rpm, and the fermented oil was fermented at 4000 rpm After centrifugation for 20 minutes, filtration was performed to obtain a fermented marine oil. The fatty acid content after fermentation was measured, and the results are shown in Table 5 below.

content (%) DHA EPA Control 1 0.94 1.85 Control 2 1.95 2.56 Example 1 8.39 10.23

As shown in Table 5, when fermenting with the culture concentrate of Rissofus oryzae according to the present invention, compared to the fermentation using the microorganism itself, even when the same marine oil was fermented, the higher omega-3 fatty acid content It is possible to obtain a fermented marine oil containing (about 9 times)

<Experimental Example 4> Measurement of yield of free fatty acid production according to fermentation temperature

In order to set the optimum fermentation conditions, the temperature of fish oil collected from sardine as marine oil was set at 37 ° C, and the culture concentrate (using 60-80% ammonium sulfate) of Experimental Example 3 was added at 10 ° C and pH 5.0 To 8.0 (Phosphate buffer (50 mM)). Thereafter, the mixture was mixed at a ratio of 50 wt% of the culture concentrate to 50 wt% of marine oil, and fermented for 72 hours at the rotation speed of 350 rpm, the reaction temperature of 25, 37, and 45 캜, respectively, .

Fermentation temperature (℃) content (%) DHA EPA 25 3.95 5.41 37 8.47 10.98 45 5.46 7.54

The results showed that the highest fatty acid production yield was obtained at 37 ℃.

<Experimental Example 5> Measurement of Yield of Fatty Acid Production by Freezing Time Sonication

In order to set the optimum fermentation conditions, the temperature of fish oil collected from sardine as marine oil was set at 37 ° C, and the culture concentrate (using 60-80% ammonium sulfate) of Experimental Example 3 was added at 10 ° C and pH 5.0 To 8.0 phosphate buffer (Phosphate buffer). Thereafter, the mixture was mixed at a ratio of 50 wt% of the culture concentrate to 50 wt% of marine oil, and ultrasonication was performed using an ultrasonic processor (manufactured by Hwasin Tech, model number: POWERSONIC 250, code: 100-520-220) After sonication, 700 W (220 V, 60 Hz) was generated, and fermentation was carried out at a rotation speed of 350 rpm and a reaction temperature of 37 ° C for 72 hours, and the results are shown in Table 7 below.

Ultrasonic treatment (minute) content (%) DHA EPA 30 4.51 6.46 60 8.46 10.25 90 7.59 9.21 120 7.17 9.01

&Lt; Experimental Example 6 > Measurement of production yield of essential fatty acid by the second fermentation

In order to set the optimum fermentation conditions, the temperature of fish oil collected from sardine as marine oil was set at 37 ° C, and the culture concentrate (using 60-80% ammonium sulfate) of Experimental Example 3 was added at 10 ° C and pH 5.0 To 8.0 phosphate buffer (Phosphate buffer). Thereafter, the mixture was mixed at a ratio of 50 wt% of the culture concentrate to 50 wt% of marine oil, fermented at a rotation number of 350 rpm, reaction temperatures of 25, 37 and 45 캜 for 72 hours, And the second fermentation was carried out under the same conditions. Analysis of the samples obtained after the first fermentation and the samples obtained after the second fermentation were measured and compared with the essential unsaturated fatty acids liberated. The results are shown in Table 8 below.

content (%) Primary fermentation Secondary fermentation DHA EPA DHA EPA 8.72 10.79 10.48 13.85

As a result of the experiment, the yield of essential fatty acid production was high in the secondary fermentation.

<Experimental Example 7> Measurement of yield of essential fatty acid production according to pH

In order to set the optimum fermentation conditions, the temperature of fish oil collected from sardine as marine oil was set at 37 ° C, and the culture concentrate (using 60-80% ammonium sulfate) of Experimental Example 3 was added at 10 ° C and pH 5.0 To 8.0 phosphate buffer (Phosphate buffer). Thereafter, the mixture was mixed at a ratio of 50% by weight of marine oil to 50% by weight of the culture concentrate, and fermented at a rotation number of 350 rpm and a reaction temperature of 37 캜 for 72 hours while varying pH alone. The results are shown in Table 9 below.

Fermentation pH content (%) DHA EPA 5.0 3.28 5.86 6.0 6.42 8.72 7.0 8.72 10.79 8.0 7.34 9.16

As a result, the highest yield of essential fatty acid production was obtained at pH 7.0 of fermentation.

<Experimental Example 8> Measurement of yield of essential fatty acid production according to fermentation and emulsifier

To set the optimal fermentation conditions, the temperature of fish oil collected from sardines as marine oil was set at 37 ° C, and the culture concentrate (using 60-80% ammonium sulfate) of Experimental Example 3 was added at 10 ° C and pH 7.0 Of phosphate buffer (Phosphate buffer). Thereafter, the mixture was mixed at a ratio of 50 wt% of the culture concentrate to 50 wt% of marine oil, and the resultant mixture was subjected to physical emulsification and an emulsifier (Tween-20, 0.2%) at a rotation speed of 350 rpm, And the results are shown in Table 10 below.

content (%) Physical oil painting Emulsifier (Tween-20, 0.2%) DHA EPA DHA EPA 8.47 10.98 9.84 11.94

Experimental results showed that there was no significant difference in yield of final unsaturated fatty acid when ultrasound and blending were used without any emulsifier.

<Experimental Example 9> Measurement of yield of essential fatty acid production according to the weight ratio of enzyme solution and oil

To set the optimal fermentation conditions, the temperature of fish oil collected from sardines as marine oil was set at 37 ° C, and the culture concentrate (using 60-80% ammonium sulfate) of Experimental Example 3 was added at 10 ° C and pH 7.0 Of phosphate buffer (Phosphate buffer). Thereafter, the mixture was mixed at a ratio of 20 to 80% by weight of the culture concentrate and 80 to 20% by weight of marine oil, and fermented at a rotation speed of 350 rpm and a reaction temperature of 37 캜 for 72 hours.

Enzyme solution: Oil
(Weight ratio,%) content (%) DHA EPA 20: 80 2.87 5.28 50: 50 7.38 9.83 80: 20 5.72 7.16

As a result of the experiment, the yield of essential fatty acid production was highest at 50:50 by weight ratio of enzyme solution and oil.

<Experimental Example 10> Measurement of yield of essential fatty acid production according to fermentation time

To set the optimal fermentation conditions, the temperature of fish oil collected from sardines as marine oil was set at 37 ° C, and the culture concentrate (using 60-80% ammonium sulfate) of Experimental Example 3 was added at 10 ° C and pH 7.0 Of phosphate buffer (Phosphate buffer). Thereafter, the mixture was mixed at a ratio of 50% by weight of marine oil to 50% by weight of the culture concentrate, and fermented at a rotation speed of 350 rpm and a reaction temperature of 37 캜 for 24 to 120 hours.

Fermentation time
(hour) content (%) DHA EPA 24 5.41 8.94 48 6.97 9.83 72 7.43 10.33 96 7.21 9.92 120 7.3 10.52

The results showed that the longer the fermentation time, the higher the yield of essential fatty acid production, but the increase rate of production of essential fatty acid was not increased after 72 hours. Therefore, considering the economical aspect and efficiency of essential fatty acid production, the fermentation time is suitable for 72 hours.

<Experimental Example 11> Measurement of yield of essential fatty acid production according to fermentation RPM

To set the optimal fermentation conditions, the temperature of fish oil collected from sardines as marine oil was set at 37 ° C, and the culture concentrate (using 60-80% ammonium sulfate) of Experimental Example 3 was added at 10 ° C and pH 7.0 Of phosphate buffer (Phosphate buffer). Thereafter, the mixture was mixed at a ratio of 50% by weight of marine oil to 50% by weight of the culture concentrate and fermented at a reaction temperature of 37 ° C at 150 to 450 rpm, and the results are shown in Table 13 below.

Fermentation speed
(RPM) content (%) DHA EPA 150 3.75 5.76 250 5.97 8.42 350 7.51 10.43 450 7.12 10.01

As the fermentation rpm increased, the highest yield of essential fatty acid production was observed. However, the rate of production of essential fatty acid increased after 350 rpm. Therefore, considering the economical aspect and efficiency of essential fatty acid production, the fermentation time is preferably 350 rpm.

EXPERIMENTAL EXAMPLE 12 Measurement of DHA and EPA Concentration in Blood of Lethal Omega-3 Oil and Omega-3 Oil after Fermentation

Five week old rats were purchased and adapted to the new breeding environment for one week and fasted for one day. One ml of pre-fermentation omega-3 oil or omega-3 oil after fermentation per litter was consumed using the ZEON. The amount of blood collected was 1.5 ml per time, and the number of blood samples was limited to three times per marriage. Therefore, in order to satisfy the collection time used in the present invention, three rats were used per sample inspection. Venous blood was collected from a tube containing 5 mM EDTA and final 20 mM sodium citrate buffer, gently shaken to prevent blood clotting, and centrifuged at 5000 rpm for 5 minutes at 4 ° C to obtain a supernatant plasma solution . 0.15 ml of chloroform: methanol (1: 1 by weight) solvent mixture was added to the plasma solution (about 0.7 ml) to extract lipid components. The extracted lipids were dissolved in methanol containing 2.5% sulfuric acid and reacted at 70 ° C. for 2 hours to form a fatty acid methyl ester (FAME). These fatty acid methyl esters were analyzed by gas chromatography and fatty acid standards were purchased from NuChek Prep.

<Experimental Example 13> Measurement of antibacterial activity

To determine the antimicrobial activity of fermented oils, two strains were selected. These include Staphylococcus aureus (Staphylococcus aureus, ATCC 25923), acne causing bacteria (Propionibacterium acnes, ATCC 49145) for We used this strain are US ATCC (American Type Culture Collection) freeze to buy that dried Mueller hinton f Bros (eller in Hinton Broth: MHB) medium 2 to 3 times.

The minimum inhibition concentration (MIC) for P. acnes and S. aureus was determined by standard liquid medium dilution method. The medium was MHB. P. acnes and S. aureus were prepared by inoculating MHB medium at a concentration of 107 cfu / ml. The samples were mixed with the prepared microorganisms such that the final concentration of the sample was 2, 4, 8% in 10% DMSO using DMSO (dimethyl sulfoxide) as a solvent and cultured at 37 ° C and 150 rpm for 16 hours. As a control group, medium containing only 10% DMSO was used. To confirm the results, 0.1 ml of Agar was added to 1.5 ml of hardened MHB, and colonies were observed after incubation at 37 ° C for 24 hours.

Samples of the fermented oil were the fermentation samples prepared in Example 7, the fermentation samples using the fermentation broth at pH 7.0, the fermentation broth at the weight ratio of the fermentation concentrate and oil at 1: 1 in Experimental Example 9, A fermentation sample obtained by fermentation for 72 hours was used. Their final concentrations were added to culture tubes as described above to 2, 4, 8% and cultured for 16 hours.

As a result, as shown in FIG. 10, the fermented oils prepared according to Experimental Examples 7, 9, and 10 prepared according to the present invention commonly have antibacterial activity against S. aureus at a final concentration of 4% Activity. However, even at a final concentration of 10% of fermented omega-3 oil, there was little antimicrobial activity against P. acnes .

Experimental Example 14 Effect of omega-3 oil on hyperlipemia in high fat diabetic rats

1. Experimental Animals: C57BL / 6 male mice, 4 weeks old, were distributed from Samtaco, Seoul, Korea. Experimental diets and drinking water were fed freely. The breeding room was maintained at a temperature of 22 ± 1 ° C and a relative humidity of 55 ± 5%, and the darkness was maintained for 12 hours (07:00 to 19:00).

2. Group separation and sample administration: For the induction of hyperlipemia, 60% kcal fat diet was fed for 5 weeks after first adjusting to the general diet for 10 days. The groups were divided into two groups: normal feed group, high fat diet group, negative control group, Omaco group, purified control group, fermented oil low group, And high concentration of oil (Fermented Oil High). Samples were fed by oral administration (once / two days) and drinking water was administered to the general and high fat feed groups (Table 14: Oral doses of each group).

group Diet Volume
(Mg / g body weight) Oral administration General feed group Normal control group 4 water High fat feed group High fat control group 4 water Omako-gun High fat feed group 2 OMOKO (fish oil) ¹⁾ Jeong's control group High fat feed group 4 Refined fish oil Low concentration of fermented oil High fat feed group 2 Fermented oil High concentration of fermentation oil High fat feed group 4 Fermented oil 1) OMARKO soft capsules: specialty medicines, KNIL PHARMACEUTICAL CO., LTD.

3. Experimental formula: The experimental diet was 60% kcal fat diet (Open Source Diets, Research Diets, Inc., New Jersey, USA) and the composition was as shown in Table 15 (G / kg diet)).

ingredient Weight (mg) Calories (kcal) Casein (80 mesh) 200 800 L-cystine 3 12 Corn starch 0 0 Maltodextrin 10 125 500 Sucrose 68.8 275.2 Cellulose (BW200) 50 0 Soybean oil 25 225 Fat 245 2205 Mineral Mix (S10026) 10 0 Dicalcium phosphate 13 0 Calcium carbonate 5.5 0 Potassium citrate (monohydrate) 16.5 0 Vitamin Mix (V10001) 10 40 Hydrogen choline tartrate 2 0 Dye (FD & C blue dye # 1) 0.05 0 Sum 773.85 4057.2

4. Sample collection: Body weight was measured once a week and dietary intake was measured twice a week during the experiment. The animals were fasted for 12 hours before sacrifice, anesthetized with diethyl ether, and then bled into the orbital vein with a Pasteur pipette. The blood was collected and the liver, kidney, epididymis, abdominal fat and epididymal fat were excised and sterilized at 0.9% After washing with sodium chloride solution, the water was removed and weighed and stored frozen at -80 ° C until analysis.

5. Serum lipid analysis: The concentration of triglyceride (AM 157S-K), total cholesterol (AM 202-K) and HDL cholesterol (AM 203-K) in the serum was measured using an enzyme reagent assay kit Lt; / RTI &gt; (ASAN Pharmaceutical). Serum triglyceride concentration was measured at 500 nm using a spectrophotometer (JENWAY 7315 Spectrophotometer, product of Bibby Scientific Ltd., Stone, England) at a concentration of total cholesterol and HDL cholesterol at 550 nm.

Statistical analysis: All results were expressed as mean and standard deviation, measured by 3 repetitions. Statistical analysis was performed using the SPSS program. The ANOVA (Scheffe, p <0.05) test was performed to verify the significance between the experimental groups.

Experiment result

1. Weight and weight gain

The changes in body weight and body weight gain of the experimental animals according to the experimental diets are shown in Table 16. After 4 weeks of feeding omega 3 oil after inducing hyperlipidemia by feeding the high fat diet for 5 weeks, weight gain was significantly decreased compared with the high fat diet group fed distilled water. In particular, The amount of increase was 3.40g, which is the least weight increase.

group Initial weight (g) Final weight (g) Weight gain (g / day) It's a highland system. 32.14 1.51 ^a 39.27 ± 1.23 ^a 7.13 ± 1.01 ^a Omako-gun 32.69 ± 1.58 ^a 37.89 ± 1.62 ^a 5.20 ± 0.80 ^b Jeong's control group 33.15 + 1.65 ^a 38.34 ± 1.99 ^a 5.20 ± 0.77 ^b Low concentration of fermented oil 33.08 + - 1.83 ^a 39.06 ± 1.64 ^a 5.98 + 0.99 ^ab High concentration of fermentation oil 33.19 ± 1.87 ^a 36.59 ± 1.51 ^a 3.40 ± 0.61 ^c

2. Lipid content in serum

The lipid content in mouse serum fed with omega-3 oil for 4 weeks was measured and is shown in Table 17. Fermented oil low concentration group, fermented oil high concentration group, and Omako group were significantly lower than those of high fat diets. The total cholesterol content was 155.0㎎ / ㎗ in the fermented oil high concentration group, which was significantly lower than that in the high fat feed group.

group Triglyceride
(Mg / dl) Total cholesterol
(Mg / dl) HDL cholesterol
(Mg / dl) It's a highland system. 110.3 + - 10.4 ^a 184.3 ± 19.6 ^a 101.3 ± 6.3 ^a Omako-gun 91.9 ± 5.4 ^b 160.6 ± 18.7 ^ab 93.5 ± 5.6 ^a Jeong's control group 100.7 ± 8.8 ^ab 168.4 ± 8.7 ^ab 103.0 + 4.2 ^a Low concentration of fermented oil 92.3 ± 8.7 ^b 164.1 ± 13.6 ^ab 95.2 + 4.2 ^a High concentration of fermentation oil 91.6 ± 7.5 ^b 155.0 ± 19.8 ^b 94.3 ± 11.3 ^a

While the invention has been shown and described with reference to certain exemplary embodiments thereof, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention as defined by the appended claims.

Claims (19)

Wherein the culture concentrate is obtained by fermenting a marine oil selected from the group consisting of fish oil, bird oil extracted from algae and a mixture thereof with a culture concentrate of Rizophus genus microorganism, In an amount of 10% by volume relative to the volume of the culture medium, culturing under culture conditions of pH 5.5 to 6.5 and 25 to 35 ° C under aerobic conditions, and then centrifuged at 800 to 1200 rpm at 1 to 10 ° C to obtain a supernatant. Wherein the fermented marine oil is a precipitate obtained by salting out with ammonium. delete The method according to claim 1,
Wherein the culture medium is PDB (Potato Dextrose Broth) containing 2 to 10% by weight of potato starch, 0.5 to 3% by weight of dextrose, and water as a remaining amount. The method of claim 3,
Wherein the PDB further comprises 0.1 to 10% by weight of oil based on the total weight of the PDB. The method according to claim 1,
Wherein the fermentation marine oil has a culture time of 80 to 150 hours under the culture conditions. The method according to claim 1,
Wherein the fermentation of the marine oil is carried out by adding a culture concentrate of the microorganism of Rizopus as a residual amount to 10 to 80% by weight of the marine oil and emulsifying the microorganism in a concentration within the range of 25 to 50 캜 Wherein the fermentation is performed by fermenting the fermented marine oil after fermentation under fermentation conditions including temperature. The method according to claim 1,
Wherein the microorganism in Rizophus is Rhizopus oryzae. A method of fermenting a marine oil to produce a fermented marine oil,
(1) adding a culture concentrate of Rizophus genus microorganism as a remainder to 10 to 80% by weight of a marine oil selected from the group consisting of fish oil, algae oil extracted from algae and a mixture thereof, and mixing to obtain a mixture, Wherein the culture concentrate is inoculated with the microorganism of the genus Rizophus in an amount of 10% by volume relative to the culture volume and cultured under aerobic conditions, pH 5.5 to 6.5, 25 to 35 ° C, Lt; RTI ID = 0.0 &gt; C &lt; / RTI &gt; at 800 to 1200 rpm, the precipitate being obtained by salting out the supernatant with ammonium sulfate;
(2) emulsifying the mixture obtained in said mixing step, wherein said emulsifying is carried out by stirring the mixture obtained in said mixing step for 5 to 20 minutes with a blender and then ultrasonifying;
(3) After the emulsifying step, the emulsified mixture is agitated in a range of 50 to 500 rpm for 24 to 120 hours under a fermentation condition including a pH within a range of 5.0 to 8.0 and a temperature within a range of 25 to 45 DEG C A fermentation step for fermentation; And
(4) a purification step of purifying the fermentation product obtained in the fermentation step;
&Lt; / RTI &gt; wherein the fermented marine oil is in the form of a mixture. delete 9. The method of claim 8,
Wherein the purification in the purification step is carried out by separating the oil layer from the aqueous layer by centrifugation of the fermentation product obtained in the fermentation step and filtering the oil layer. delete 9. The method of claim 8,
Wherein the culture medium is PDB (Potato Dextrose Broth) containing 2 to 10% by weight of potato starch, 0.5 to 3% by weight of dextrose, and water as a remaining amount. 9. The method of claim 8,
Wherein the culturing time is within a range of 80 to 150 hours under the culturing condition. 9. The method of claim 8,
Wherein the microorganism in Rizophus is Rhizopus oryzae. delete delete delete 7. An antimicrobial composition comprising fermented marine oil according to any one of claims 1 to 7 as an active ingredient.
delete

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